1,25(OH)_2D_3对体外培养大鼠成骨细胞RANKL/OPG mRNA表达的影响  

Effect of 1,25(OH)2D3 on the expression of RANKL and osteoprotegerin mRNA in primary rat osteoblasts

作　　者：吴小涛[1] 王海军[1] 宋萍[1] 

机构地区：[1]东南大学附属中大医院骨科,江苏南京210009

出　　处：《东南大学学报（医学版）》2007年第1期32-35,共4页Journal of Southeast University(Medical Science Edition)

摘　　要：目的:观察1,25(OH)2D3对体外培养的大鼠成骨细胞RANKL/OPG mRNA表达的影响,探讨其促进骨吸收的作用机制。方法:体外分离培养大鼠成骨细胞,分别观察不同浓度1,25(OH)2D3及用1,25(OH)2D3处理不同时间对RANKL/OPGmRNA表达的影响,RANKL/OPG mRNA表达采用半定量逆转录聚合酶链反应(RT-PCR)方法测定。结果:1,25(OH)2D3呈时间和剂量依赖性地刺激大鼠成骨细胞RANKL mRNA的表达,而抑制OPG mRNA的表达,使RANKL/OPG值增高。结论:1,25(OH)2D3通过调节成骨细胞RANKL/OPG的基因表达,从而发挥促进破骨细胞介导的骨吸收作用。Objective To observe the effect of 1,25 （OH）2D3on the expression of RAN KL and osteoprotegerin （OPG） mRNA in primary rat osteoblasts. Methods Primary osteoblastic cells were obtained from the calvaria of newborn SD rats by digestion with 0. 1% collagenase and cultured in PRMI 1640 medium supplemented with 15% fetal bovine serum. Cells were treated with 1,25 （OH）2D3 according to the experimental design. Total cellular RNA was isolated with Trizol. Semiquantitative reverse transcription RCR method was used to screen the level of RANKL and OPG mRNA. Results RANKL mRNA expression in the calvarial osteoblasts was elevated dose-dependently following 1,25 （OH） 2 D3 treatment and reached its peak level at a concentration of 10^-8 mol·L^-1 （ P 〈 0.01 ）. 1,25 （ OH ） 2 D3 inhibited OPG mRNA levels by 60% at a concentration 10^-7mol·L^-1 after 48 h（P 〈0.01 ）. 1,25（OH）2D3（ 10^-7mol·L^-1 ）timedependently resulted in a up h（ P 〈 0. 01 ）. Conclusions regulation of RANKL mRNA levels and a downregulation of OPG mRNA peaking both at 48 1,25 （OH）2D3enhanced RANKL and inhibited OPG mRNA expression of primary rat osteoblasts in a time-and dose-dependent fashion. By altering the relative expression of OPG and RANKL mRNA, 1,25 （OH）2D3 might stimulate the osteoclastic bone resorption.

关 键 词：1 25(OH)2D3 成骨细胞 核因子κB受体活化子配体 保骨素 

分 类 号：R589.5[医药卫生—内分泌]